RT Journal Article
SR Electronic
T1 Production, Characterization, and Uses of Monoclonal Antibodies against Recombinant Nucleoprotein of Elk Coronavirus
JF Clinical and Diagnostic Laboratory Immunology
JO CLIN. DIAGN. LAB. IMMUNOL.
FD American Society for Microbiology
SP 341
OP 344
DO 10.1128/CDLI.6.3.341-344.1999
VO 6
IS 3
A1 Daginakatte, Girish C.
A1 Chard-Bergstrom, Cindy
A1 Andrews, Gordon A.
A1 Kapil, Sanjay
YR 1999
UL http://cvi.asm.org/content/6/3/341.abstract
AB This is the first report of the production of monoclonal antibodies against elk coronavirus. The nucleoprotein gene of elk coronavirus was amplified by PCR and was cloned and expressed in a prokaryotic expression vector. Recombinant nucleocapsid protein was used to immunize mice for the production of hybridomas. Twelve hybridomas that produced monoclonal antibodies against the nucleocapsid protein of elk coronavirus were selected by an indirect fluorescent-antibody test, an enzyme-linked immunosorbent assay, and a Western blot assay. Ten of the monoclonal antibodies were of the immunoglobulin G1 (IgG1) isotype, one was IgG2a, and one was IgM. All had kappa light chains. By immunohistochemistry four monoclonal antibodies detected bovine coronavirus and elk coronavirus in formalin-fixed intestinal tissues. Antinucleoprotein monoclonal antibodies were found to be better at ruminant coronavirus detection than the anti-spike protein monoclonal antibodies. Because nucleoprotein is a more abundant antigen than spike protein in infected cells, this was not an unexpected finding.